中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (6): 901-905.doi: 10.3969/j.issn.2095-4344.1579

• 膜生物材料 membrane biomaterials • 上一篇    下一篇

基于静电纺织技术构建血液生物膜的初探及生物相容性评价

孙 宇1，2，3，邹 强1，2，3，李轩泽1，2，3，吴展羽1，2，杨 龙2，王建吉2，刘 琴4，马敏先4，叶 川1，2，3   

  1. 1贵州医科大学，贵州省贵阳市 550004；2贵州医科大学附属医院骨科，贵州省贵阳市 550004；3贵州医科大学组织工程与干细胞实验中心，贵州省贵阳市 550004；4贵州医科大学附属口腔医院修复科，贵州省贵阳市 550004
  • 收稿日期:2018-09-13 出版日期:2019-02-28 发布日期:2019-02-28
  • 通讯作者: 叶川，主任医师，贵州医科大学附属医院骨科，贵州省贵阳市 550004
  • 作者简介:孙宇，女，1991年生，黑龙江省鹤岗市人，汉族，贵州医科大学在读硕士，医师，主要从事组织工程生物材料的研发工作。
  • 基金资助:

    国家自然科学基金资助项目(81360232)，项目负责人：叶川；贵阳市科技局创新团队资助项目(20175-17)，项目负责人：叶川

Construction and biocompatibility of blood biofilms based on electrospinning technology

Sun Yu1, 2, 3, Zou Qiang1, 2, 3, Li Xuanze1, 2, 3, Wu Zhanyu1, 2, Yang Long2, Wang Jianji2, Liu Qin4, Ma Minxian4, Ye Chuan1, 2, 3   

  1. 1Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 2Department of Orthopedics, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 3National and Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique, Guiyang 550004, Guizhou Province, China; 4Department of Prosthodontics, Affiliated Stomatology Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Received:2018-09-13 Online:2019-02-28 Published:2019-02-28
  • Contact: Ye Chuan, Chief physician, Guizhou Medical University, Guiyang 550004, Guizhou Province, China; Department of Orthopedics, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China; National and Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique, Guiyang 550004, Guizhou Province, China
  • About author:Sun Yu, Master candidate, Physician, Guizhou Medical University, Guiyang 550004, Guizhou Province, China; Department of Orthopedics, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China; National and Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique, Guiyang 550004, Guizhou Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81360232 (to YC); the Science and Technology Innovation Team Funded Project of Guiyang Municipal Science and Technology Department, No. 20175-17 (to YC)

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摘要：

文章快速阅读：

 

文题释义：
静电纺织技术：是一种特殊的纤维制造工艺，将聚合物溶液或熔体在强电场中进行喷射纺丝；在电场作用下，针头处的液滴会由球形变为圆锥形(即“泰勒锥”)，并从圆锥尖端延展得到纤维细丝，这种方式可生产出纳米级纤维结构的支架材料。
纤维蛋白原：是一种具有凝血功能的蛋白质，在组织工程领域可作为支架材料，该材料可促进细胞的黏附、增殖，指导细胞行为，具有良好的生物相容性。
低温喷雾干燥技术：是物料干燥的方法之一，可直接使溶液、乳浊液干燥成粉状，优点为：干燥过程非常迅速，可直接干燥成粉末，低温不影响生物材料的活性。
 
 
背景：已有研究表明纤维蛋白原可作为组织工程支架材料，若将血液中的纤维蛋白原提纯再利用，可制作出无免疫原性、生物相容性良好的支架材料。
目的：探索静电纺丝技术制备血液生物膜的方法，评价支架的生物相容性。
方法：将新鲜猪血液经分离、离心、纯化等步骤提纯出纤维蛋白原，分别经真空冷冻干燥机(A组)、低温喷雾干燥机(B组)、56 ℃烘箱干燥(C组)处理，采用静电纺织技术制作成纤维薄膜支架材料，检测3组支架材料的接触角，采用扫描电镜观察支架材料的三维结构。将3组电纺支架材料分别与骨髓间充质干细胞共培养，培养7 d后，扫描电镜观察细胞生长情况；培养1-4 d，Alamar Blue试剂盒检测B组支架材料表面细胞增殖。

结果与结论：①扫描电镜显示，A组可见纤维结构，纤维粗细不均并有大量液滴状结构；B组可见排列有序的多层纤维结构，纤维直径、孔径大小接近一致，仅见少量液滴状结构；C组未见纤维结构，为大小不等的液滴状结构；②A-C组支架材料的接触角分别为(82±3)°、(67±5)°、(80±3)°，3组材料接触角都< 90°，表明亲水性良好；③共培养7 d 后，骨髓间充质干细胞均可黏附于3组支架材料上，其中B组材料表面的细胞分布较为均匀，细胞黏附生长于纤维表面，细胞核形态相对规则；A、C组可见少量细胞黏附生长于支架上，C组细胞量最少，细胞核形态不规则，两组均未见明显纤维结构；④Alamar Blue实验结果显示，随着培养时间的延长，B组支架材料表面的细胞呈增殖趋势，生长状态良好；⑤结果表明，采用静电纺织技术可制备出血液纤维蛋白原生物膜，其具有良好的生物相容性。

ORCID: 0000-0002-5247-6858(孙宇)

关键词: 维蛋白原, 血液生物膜, 静电纺织, 纤维蛋白原, 细胞增殖趋势, 血液纤维蛋白原生物膜

Abstract:

BACKGROUND: Previous studies have shown that fibrinogen can serve as tissue engineering scaffold material. Therefore, the reuse of fibrinogen in the autologous blood can make the scaffold material which has non-immunogenicity and good biocompatibility.

OBJECTIVE: To explore the method of preparing blood biofilm using the electro-spinning technology and to evaluate the biocompatibility of the fibrinogen scaffold.
METHODS: Fibrinogen was purified out of the fresh blood after the procedures of separation, centrifugation, and purification, and then subjected to vacuum freeze drying (group A), low temperature spray drying (group B), and oven drying at 56 oC (group C). It was finally made into fiber film using the electrospinning technology. Contact angle of the three sets of scaffold materials was detected, and the three-dimensional structure of the scaffolds was observed under scanning electron microscope. Bone marrow mesenchymal stem cells were co-cultured with the three sets of scaffolds in vitro. Cell growth was observed using scanning electron microscope at 7 days of culture, and cell proliferation in the group B was detected using Alamar Blue kit at 1-4 days of culture.

RESULTS AND CONCLUSION: (1) Under the scanning electron microscope, fiber structure with uneven fiber thickness was observed, and a lot of droplet-like structures were also visible in the group A; the ordered multi-layer fiber structure with similar fiber diameter and pore size was observed in the group B, and only a small amount of droplet-like structures were detected; in the group C, there was no fiber structure, but the droplet-like structures of different sizes. (2) The contact angle of the scaffold material was (82±3)o in the group A, (67±5)o in the group B, and (80±3)o in the group C. (3) After 7 days of co-culture, the bone marrow mesenchymal stem cells adhered to the scaffold materials in the three groups. The cells on the surface of group B were evenly distributed, which were adherent to the fiber surface and had relative regular nuclei. Only a small amount of cells grew onto the scaffolds in the groups A and C, especially in the group C, in which the cells had irregular nuclei and no fiber structure was observed. (4) The cells in the group B exhibited an increasing proliferation tendency as shown by the results of Alamar Blue experiment, and the cell growth was in good status. All these findings indicate that it is feasible to prepare the blood fibrinogen biofilm with good biocompatibility using the electrospinning technology. 

Key words: Fibrinogen, Biofilms, Blood;, Electrospinning, Tissue Engineering

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